2. Academic Validation
  3. Inflammatory Caspases: Activation and Cleavage of Gasdermin-D In Vitro and During Pyroptosis

Inflammatory Caspases: Activation and Cleavage of Gasdermin-D In Vitro and During Pyroptosis

  • Methods Mol Biol. 2018:1714:131-148. doi: 10.1007/978-1-4939-7519-8_9.
Yue Zhao 1,  Jianjin Shi 1,  Feng Shao 2
Affiliations

Affiliations

  • 1 National Institute of Biological Sciences, #7 Science Park Rd, Beijing, 102206, China.
  • 2 National Institute of Biological Sciences, #7 Science Park Rd, Beijing, 102206, China. [email protected].
PMID: 29177860 DOI: 10.1007/978-1-4939-7519-8_9
Abstract

Gasdermin-D (also known as GSDMD), the newly identified executioner of pyroptotic cell death, is cleaved by activated Caspase-1 downstream of canonical inflammasome activation or caspase-4, 5, and 11 upon their ligation and activation by cytosolic LPS. Upon a single cleavage between the two domains in Gasdermin-D, the N-terminal domain binds to membrane lipids and lyses cells by forming pores of an inner diameter of 10-14 nm within the membrane. The inter-domain cleavage of Gasdermin-D is a reliable marker for the activation of inflammatory caspases and cell Pyroptosis. Here, we describe the methods for examining Gasdermin-D cleavage by activated inflammatory caspases in vitro and upon inflammasome activation in vivo.

Keywords

GSDMD; Gasdermin-D; Inflammasome; Inflammatory caspases; LPS; Pyroptosis; Recombinant proteins.

Figures